Giesbrecht M, McCarthy M, Elliott M L, Ong K L
Texas A&M AgriLife Extension Service, College Station, TX.
University of Florida-IFAS, Fort Lauderdale Research and Extension Center.
Plant Dis. 2013 Nov;97(11):1511. doi: 10.1094/PDIS-05-13-0488-PDN.
Fusarium wilt of palms occurs worldwide, caused by different Fusarium oxysporum ff. spp. including F. oxysporum f. sp. elaeidis, F. oxysporum f. sp. canariensis, and F. oxysporum f. sp. albedinis (3). Prior to 2010, F. oxysporum f. sp. canariensis was the only palm infecting species known to occur in the United States. In 2010, isolates of F. oxysporum were reported from dying Syagrus romanzoffiana and Washingtonia robusta in Florida. Based on morphological and molecular data, as well as the unique host species affected by the pathogen, this fungus was determined to be a new forma specialis of F. oxysporum, designated f. sp. palmarum (1). The pathogen infects foliar tissue, causing complete necrosis of the crown and leading to tree death within 2 to 3 months. In June 2012, the Texas Plant Disease Diagnostic Laboratory (TPDDL) received a plant sample from a dying W. robusta palm, exhibiting reddish-brown stripes on the petiole with chlorotic and necrotic leaves, from an established palm in the landscape from Harris County, Texas. Fungal cultures were obtained from symptomatic foliar tissue and identified as F. oxysporum based on morphology. Microconidia were oval to reniform, 1- to 2-septate, measuring 5 to 18 × 2.5 to 5 μm. Phialides were short with microconidia produced in false heads. Macroconidia were curved and slender with a foot-shaped basal cell, usually 3-septate, and 22 to 37 × 2.5 to 5 μm. Chlamydospores were roundish and ranged from 7 to 13 μm in diameter. Fungal colonies had white to purple mycelia when grown on potato dextrose agar. DNA from a single spore culture was extracted, amplified by PCR using primers corresponding to a segment of the translation elongation factor 1α (EF-1α) gene, and the PCR product sequenced (2). Using the sequence alignment tool (BLASTn) in GenBank, the TPDDL's sequence (GenBank Accession No. KC897693) was aligned with EF-1α regions from F. oxysporum f. sp. palmarum isolates previously entered into the database ([1]; accessions GQ154455[=NRRL53544] and GQ154456[=NRRL46589]), revealing 100% homology between the isolates. Based on host source and sequence similarity, the fungus was tentatively identified as F. oxysporum f. sp. palmarum. Pathogenicity tests were performed on three leaf seedlings of W. robusta and W. filifera. Fifteen plants of each species were inoculated with the suspect isolate (designated KB2012) and 10 control plants were mock-inoculated as described by (1). Plants were grown in a greenhouse for 8 weeks post-inoculation. During this time, 83% of inoculated plants developed foliar lesions and died or severely declined, and all control plants remained healthy. F. oxysporum was recovered in culture from 100% of the symptomatic plants. DNA was extracted from fungal cultures, and EF-1α was amplified by PCR and sequenced, as described above. The amplicon was determined to share 100% homology with known F. oxysporum f. sp. palmarum isolates, confirming this fungus as the cause of disease in W. robusta. This is the first report of this pathogen in Texas, as well as the first report outside of Florida. This is also the first documentation of W. filifera as a host of this pathogen. References: (1) M. L. Elliott et al. Plant Dis. 94:31, 2010. (2) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (3) G. W. Simone. Pages 17-19 in: Compendium of Ornamental Palm Diseases and Disorders, M. L. Elliott et al., eds. The American Phytopathological Society, St. Paul, MN, 2004.
棕榈枯萎病在全球范围内均有发生,由不同的尖孢镰刀菌专化型引起,包括尖孢镰刀菌油棕专化型、尖孢镰刀菌加那利椰枣专化型和尖孢镰刀菌海枣专化型(3)。2010年之前,尖孢镰刀菌加那利椰枣专化型是已知在美国出现的唯一能感染棕榈的菌种。2010年,在佛罗里达州,从濒死的皇后葵和华盛顿葵上分离出了尖孢镰刀菌菌株。基于形态学和分子数据,以及受该病原菌影响的独特寄主物种,这种真菌被确定为尖孢镰刀菌的一个新专化型,命名为棕榈专化型(1)。该病原菌感染叶片组织,导致树冠完全坏死,并在2至3个月内导致树木死亡。2012年6月,得克萨斯州植物病害诊断实验室(TPDDL)从得克萨斯州哈里斯县一处园林中一棵濒死的华盛顿葵上收到一份植物样本,该植株叶柄上有红棕色条纹,叶片出现褪绿和坏死症状。从有症状的叶片组织中获得真菌培养物,并根据形态学鉴定为尖孢镰刀菌。分生孢子呈椭圆形至肾形,具1至2个隔膜,大小为5至18×2.5至5μm。瓶梗短,分生孢子呈假头状着生。大型分生孢子弯曲且细长,基部细胞呈足形,通常具3个隔膜,大小为22至37×2.5至5μm。厚垣孢子呈圆形,直径为7至13μm。真菌菌落生长在马铃薯葡萄糖琼脂上时,菌丝体为白色至紫色。从单孢培养物中提取DNA,使用对应于翻译延伸因子1α(EF - 1α)基因片段的引物进行PCR扩增,并对PCR产物进行测序(2)。使用GenBank中的序列比对工具(BLASTn),将TPDDL的序列(GenBank登录号KC897693)与之前录入数据库的尖孢镰刀菌棕榈专化型分离株的EF - 1α区域进行比对([1];登录号GQ154455[=NRRL53544]和GQ154456[=NRRL46589]),结果显示分离株之间具有100%的同源性。基于寄主来源和序列相似性,该真菌初步鉴定为尖孢镰刀菌棕榈专化型。对三株华盛顿葵和三株加州蒲葵的叶片幼苗进行了致病性测试。按照(1)中所述方法,对每个物种的15株植株接种可疑分离株(命名为KB2012),并对10株对照植株进行 mock接种。接种后,将植株在温室中培养8周。在此期间,83%的接种植株出现叶片病斑并死亡或严重衰弱,而所有对照植株均保持健康。从100%的有症状植株中培养出了尖孢镰刀菌。如上文所述,从真菌培养物中提取DNA,通过PCR扩增EF - 1α并进行测序。扩增产物被确定与已知的尖孢镰刀菌棕榈专化型分离株具有100%的同源性,证实该真菌是华盛顿葵病害的病原菌。这是该病原菌在得克萨斯州的首次报道,也是佛罗里达州以外的首次报道。这也是加州蒲葵作为该病原菌寄主的首次记录。参考文献:(1)M. L. Elliott等人,《植物病害》94:31,2010年。(2)D. M. Geiser等人,《欧洲植物病理学杂志》110:473,2004年。(3)G. W. Simone,载于:《观赏棕榈病害与失调汇编》,M. L. Elliott等人编,美国植物病理学会,明尼苏达州圣保罗,2004年,第17 - 19页。